Autonomous Detection of Aerosolized Biological Agents by Multiplexed Immunoassay with Polymerase Chain Reaction Confirmation

Benjamin J. Hindson,† Mary T. McBride,† Anthony J. Makarewicz,† Bruce D. Henderer,†
Ujwal S. Setlur,† Sally M. Smith,† Dora M. Gutierrez,† Thomas R. Metz,† Shanavaz L. Nasarabadi,† Kodumudi S. Venkateswaran,† Stephen W. Farrow,‡ Bill W. Colston, Jr.,† and John M. Dzenitis*,†

The autonomous pathogen detection system (APDS) is an automated, podium-sized instrument that continuously monitors the air for biological threat agents (bacteria, viruses, and toxins). The system has been developed to warn of a biological attack in critical or high-traffic facilities and at special events. The APDS performs continuous aerosol collection, sample preparation, and detection using multiplexed immunoassay followed by confirmatory PCR using real-time TaqMan assays. We have integrated completely reusable flow-through devices that perform DNA extraction and PCR amplification. The fully inte- grated system was challenged with aerosolized Bacillus anthracis, Yersinia pestis, Bacillus globigii, and botulinum toxoid. By coupling highly selective antibody- and DNA-based assays, the probability of an APDS reporting a false positive is extremely low.

Aerosol monitoring systems that test for the presence of biological agents are in use throughout the United States.1 Their purpose is to detect a biological agent release so that early public health action may be taken. The impact of a biological agent attack can be reduced if the appropriate responses are executed with minimal delay.1 For example, at the 2002 Winter Olympics in Salt Lake City, aerosol samples from dry filter collectors were periodi- cally transported to a centralized field laboratory where they were screened for the presence of pathogen specific nucleic acid sequences.

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